Comparative studies of the preparation of immunoliposomes with the use of two bifunctional coupling agents and investigation of in vitro immunoliposome-target cell binding by cytofluorometry and electron microscopy

The two coupling agents SPDP (N-succinimidyl-3-(2-pyridyldithio)propionate) and SATA (N-succinimidyl-S-acetylthioacetate) were compared in their efficiency and feasibility to couple monoclonal antibodies (Abs) via thioether linkage to liposomes functionalized by various lipophilic maleimide compounds like Full-size image methyl ester (MP-PL), N-(3-maleimidopropionyl)phosphatidylethanolamide (MP-PE), Full-size image methyl ester (EMC-PL), and N-(6-maleimidocaproyl)phosphatidylethanolamine (EMC-PE). The composition of the liposomes was soy phosphatidylcholine (SPC), cholesterol, maleimide compounds and -tocopherol (1:0.2:0.02:0.01, mol parts), plus N4-oleylcytosine arabinoside (NOAC) as cytostatic prodrug (0.2 mol parts) and a new, lipophilic and highly fluorescent dye N,N′-bis(1-hexylheptyl)-3,4:9,10-perylenebis(dicarboximide) (BHPD, 0.006 mol parts). From the maleimide derivatives MP-PL was the most effective in terms of preservation of the coupling activity in dependence of liposome storage. The coupling of the monoclonal A B8-24.3 (mouse IgG2b, MHC class I, anti H-2kb) and IB16-6 (rat IgG2a, anti B16 mouse melanoma) to the drug carrying liposomes was more effective and easier to accomplish with SATA as compared to SPDP. Coupling rates of 60–65% were obtained with SATA at molar ratios of 12 SATA:1 Ab:40 maleimide spacer groups on the surface of one liposome. The highest coupling rates with SPDP were obtained at the ratio of 24 SPDP:1 Ab:40 liposomal maleimide groups, with an Ab binding efficiency of only 20–25%. The optimal in vitro binding conditions to specific target cells (EL4 for B8-24.3-liposomes and B16-F10 for IB16-6-liposomes) were determined by cytofluorometric measurement of the liposomal BHPD fluorescence with SATA linked Abs. Optimal immunoliposome binding to specific epitopes on the target cells was achieved with 1–2 Ab molecules coupled to one liposome, with immunoliposome concentrations of 20–130 nM and with a small incubation volume of 0.3–0.4 ml. The specificity of the binding of B8-24.3-liposomes to EL4 target cells was visualized by scanning electron microscopy. Antibody mediated endocytic uptake of immunoliposomes could be demonstrated by transmission electron microscopy

Exploring and contextualizing public opposition to renewable electricity in the United States

This article explores public opposition to renewable power technologies in the United States. It begins by discussing the genesis of environmental ethics, or how some Americans have come to place importance on the protection of the environment and preservation of species, ecosystems, and the biosphere. As result, renewable power systems have become challenged on ethical and environmental grounds and are occasionally opposed by local communities and environmentalists. The article finds that, however, such concern may be misplaced. Renewable electricity resources have many environmental benefits compared to power stations fueled by coal, oil, natural gas, and uranium. Opposition towards renewable resources can at times obscure the true costs and risks associated with electricity use and entrench potential racial and class-based inequalities within the current energy system

IRX-2, a Novel Immunotherapeutic, Enhances Functions of Human Dendritic Cells

Background: In a recent phase II clinical trial for HNSCC patients, IRX-2, a cell-derived biologic, promoted T-cell infiltration into the tumor and prolonged overall survival. Mechanisms responsible for these IRX-2-mediated effects are unknown. We hypothesized that IRX-2 enhanced tumor antigen-(TA)-specific immunity by up-regulating functions of dendritic cells (DC). Methodology/Principal Findings: Monocyte-derived DC obtained from 18 HNSCC patients and 12 healthy donors were matured using IRX-2 or a mix of TNF-α, IL-1β and IL-6 ("conv. mix"). Multicolor flow cytometry was used to study the DC phenotype and antigen processing machinery (APM) component expression. ELISPOT and cytotoxicity assays were used to evaluate tumor-reactive cytotoxic T lymphocytes (CTL). IL-12p70 and IL-10 production by DC was measured by Luminex® and DC migration toward CCL21 was tested in transwell migration assays. IRX-2-matured DC functions were compared with those of conv. mix-matured DC. IRX-2-matured DC expressed higher levels (p<0.05) of CD11c, CD40, CCR7 as well as LMP2, TAP1, TAP2 and tapasin than conv. mix-matured DC. IRX-2-matured DC migrated significantly better towards CCL21, produced more IL-12p70 and had a higher IL12p70/IL-10 ratio than conv. mix-matured DC (p<0.05 for all). IRX-2-matured DC carried a higher density of tumor antigen-derived peptides, and CTL primed with these DC mediated higher cytotoxicity against tumor targets (p<0.05) compared to the conv. mix-matured DC. Conclusion: Excellent ability of IRX-2 to induce ex vivo DC maturation in HNSCC patients explains, in part, its clinical benefits and emphasizes its utility in ex vivo maturation of DC generated for therapy. © 2013 Schilling et al

Efectos de diferentes inmunomoduladores en la respuesta inmune a vacunas antiaftosa

La Fiebre Aftosa es la enfermedad viral del ganado que mayor impacto económicoocasiona en Argentina, no solo por los perjuicios directos sobre las especies afectadas, sinotambién por las perdidas indirectas debido al cierre de mercados de exportacion querepresentarían divisas para nuestro país. El principal elemento utilizado en la lucha contra esta enfermedad es la inmunizaciónpreventiva, pero la principal limitante de los inmunógenos en uso es la corta duracion deinmunidad que confieren. En este trabajo se utilizo el modelo murino para estudiar el efecto de tres adyuvantesdiferentes [avridine, extracto de pared celular de Mycobacteria (PCM) y lipopolisacárido de Brucella ovis(LPS)] en la respuesta a vacunas antiaftosa. Se examinó la duración deinmunidad conferida, el estado de protección viral 7 meses post-vacunación, las celulasinmunocompetentes estimuladas, involucradas en la respuesta secundaria de larga duracion ylos isotipos de IgG inducidos por los diferentes adyuvantes. La incorporación de avridine, LPS o PCM en vacunas convencionales indujo unarespuesta de anticuerpos neutralizantes especificos potente y duradera. Los índicesseroneutralizantes de animales vacunados con las nuevas formulaciones fueronsignificativamente mayores que los observados en animales inmunizados con vacunasconvencionales y confirieron proteccion por lo menos durante 7 meses. Los datos que surgen de los ensayos de reconstitución indican que losinmunomoduladores probados participan en la activación de las poblaciones celularesinvolucradas en la memoria de larga duracion, lo que resulta en una respuesta secundaria decélulas B eficiente aún en ausencia de celulas T, si bien es imprescindible la presencia delinfocitos T para la accion primaria de estos adyuvantes. Los linfocitos T se ven estimulados demanera tal que adquieren la capacidad de estimular a linfocitos B primados contra el antígenopara la producción de anticuerpos. Avridine, PCM y LPS estimulan la produccion de IgGl, 2a y 2b. lo cual se encontraríarelacionado con la proteccion conferida por las vacunas formuladas con estosinmunomoduladores. No se detectaron reacciones adversas salvo en el caso del uso de vehículo oleoso con LPS y PCM que produjo una sensibilización de las células esplenicas por la cual no fue posibleextraer células viables en estos caso 180 dias post-vacunación. Los efectos de avridine, LPS y PCM en la respuesta inmune a vacunas antiaftosaobservadas en el modelo murino indican la posibilidad de su inclusión en vacunas para serensayadas en bovinos.Fil: Berinstein, Analía. Universidad de Buenos Aires. Facultad de Ciencias Exactas y Naturales; Argentina

Functional ectodomain of the hemagglutinin-neuraminidase protein is expressed in transgenic tobacco cells as a candidate vaccine against Newcastle disease virus.

Recently, the use of plants for the production of recombinant proteins has been well demonstrated with promising outcomes. In this study, an efficient Nicotiana tabacum L. cv. Bright Yellow 2 (BY-2) cells system expressing the ectodomain of hemagglutinin-neuraminidase (eHN) protein from Newcastle disease virus (NDV) strain AF2240 was established. Transgenic tobacco BY-2 cell cultures expressing the immunogenic eHN protein were generated and the translation efficiency of eHN protein was enhanced using the 5′-untranslated region of Nicotiana tabacum alcohol dehydrogenase gene (NtADH 5′-UTR) under the control of strong cauliflower mosaic virus (CaMV 35S) promoter. Transgenic lines verified by real-time PCR showed high level of eHN mRNA transcripts and immunoblotting confirmed the presence of 66 kD eHN protein. The eHN protein was stably produced in an average of 0.2–0.4 % total soluble protein. Green fluorescent protein-tagged eHN protein was expressed and localized at the cytosol of BY-2 cell. All mice receiving purified eHN protein from transgenic tobacco BY-2 cells produced specific anti-NDV antibodies. We concluded that plant made eHN elicit immune response and can serve as candidate vaccine against NDV

Projects on the geometry of perception and cognition

Thesis (S.M. in Art, Culture and Technology)--Massachusetts Institute of Technology, Dept. of Architecture, 2012.This electronic version was submitted by the student author. The certified thesis is available in the Institute Archives and Special Collections.Cataloged from student-submitted PDF version of thesis.Includes bibliographical references (p. [98]-[102]).The projects presented in this thesis, which include performance, photography, and sculpture, investigate perception and cognition through the study and reconfiguration of content drawn from philosophy, cognitive science, and linguistics. I suspect that the language that we use to communicate about perception may be faulty. Within this critical perspective, the projects are propositions in response to the question: What is the form of perception/cognition? Underlying the projects is a fundamental philosophical question: Why do we have conscious experience? In philosophy these are referred to, respectively, as the hard and easy problems of consciousness. I investigate the linguistic structures of 'language' and 'parole' in a related attempt to understand the function of language, first independently, and then within a cognitive framework. The experiments begin with words such as 'definition', 'vision', 'perception' and represent systems defined by these signifies using objects, actions, and images. Reconfiguring the words into tangible experiments allows the nature of the phenomenon to be examined outside of the limitations of linguistic description. Ideally, the incongruity that might exist between the words and the experiences of perception and cognition can be uncovered through this process.by Sofia Rebeca Berinstein.S.M.in Art, Culture and Technolog

Living with Precarious Legal Status in Canada: Implications for the Well-Being of Children and Families

This study focused on the effects of precarious status on the well-being of fifteen participants with particular attention to their attempts to claim services, their feelings of belonging and sense of social support, and the effects of parents’ status on children. It investigates ways in which the status of one family member can affect the well-being of the entire family. Those who had children reported that the family’s status disadvantaged their children, whether they were Canadian or foreign-born, as parents’ status was used to justify denying children rights to which they are entitled by international, national, and provincial laws. The paper challenges approaches to citizenship and immigration status that fail to consider the implications of legal status for a person’s primary social units and networks.Cette étude examine les conséquences du statut précaire sur le bien-être de 15 participants, en se penchant tout particulièrement sur leurs efforts pour revendiquer l’accès aux services, leurs sentiments d’appartenance et de soutien social, ainsi que les répercussions du statut des parents sur leurs enfants. Elle examine les différentes façons par lesquelles le statut d’un membre de la famille peut affecter le bien-être de la famille toute entière. Ceux ayant des enfants ont rapporté que ces derniers, qu’ils soient nés au Canada ou à l’étranger, avaient été défavorisés par le statut de la famille, étant donné que le statut des parents était employé pour justifier le déni aux enfants de droits qui étaient les leurs en droit international et selon les lois nationales et provinciales. L’article remet en question les façons d’aborder la question de statut de citoyenneté et d’immigrant qui ne prennent pas en ligne de compte les conséquences du statut juridique sur les unités sociales de base et les réseaux sociaux pour chaque personne

Rituximab in B-Cell Hematologic Malignancies: A Review of 20 Years of Clinical Experience

Rituximab is a human/murine, chimeric anti-CD20 monoclonal antibody with established efficacy, and a favorable and well-defined safety profile in patients with various CD20-expressing lymphoid malignancies, including indolent and aggressive forms of B-cell non-Hodgkin lymphoma. Since its first approval 20 years ago, intravenously administered rituximab has revolutionized the treatment of B-cell malignancies and has become a standard component of care for follicular lymphoma, diffuse large B-cell lymphoma, chronic lymphocytic leukemia, and mantle cell lymphoma. For all of these diseases, clinical trials have demonstrated that rituximab not only prolongs the time to disease progression but also extends overall survival. Efficacy benefits have also been shown in patients with marginal zone lymphoma and in more aggressive diseases such as Burkitt lymphoma. Although the proven clinical efficacy and success of rituximab has led to the development of other anti-CD20 monoclonal antibodies in recent years (e.g., obinutuzumab, ofatumumab, veltuzumab, and ocrelizumab), rituximab is likely to maintain a position within the therapeutic armamentarium because it is well established with a long history of successful clinical use. Furthermore, a subcutaneous formulation of the drug has been approved both in the EU and in the USA for the treatment of B-cell malignancies. Using the wealth of data published on rituximab during the last two decades, we review the preclinical development of rituximab and the clinical experience gained in the treatment of hematologic B-cell malignancies, with a focus on the well-established intravenous route of administration. This article is a companion paper to A. Davies, et al., which is also published in this issue

Therapeutic vaccines and cancer: focus on DPX-0907

In an attempt to significantly enhance immunogenicity of peptide cancer vaccines, we developed a novel non-emulsion depot-forming vaccine platform called DepoVax™ (DPX). Human leukocyte antigen (HLA)-A2 restricted peptides naturally presented by cancer cells were used as antigens to create a therapeutic cancer vaccine, DPX-0907. In a phase I clinical study, the safety and immune-activating potential of DPX-0907 in advanced-stage breast, ovarian, and prostate cancer patients were examined, following encouraging results in HLA-A2 transgenic mice. The DPX-0907 vaccine was shown to be safe and well tolerated, with injection-site reactions being the most commonly reported adverse event. Vaccinated cancer patients exhibited a 61% immune response rate, with higher response rates in the breast and ovarian cancer patient cohorts. In keeping with the higher immune efficacy of this vaccine platform, antigen-specific responses were detected in 73% of immune responders after just one vaccination. In 83% of responders, peptide-specific T-cells were detected at two or more time points post-vaccination, with 64% of these patients showing evidence of immune persistence. Immune monitoring also demonstrated the generation of antigen-specific T-cell memory, with the ability to secrete multiple type 1 cytokines. The novel DPX formulation promotes multifunctional effector/memory responses to peptide-based tumor-associated antigens. The data support the capacity of DPX-0907 to elicit type-1 biased immune responses, warranting further clinical development of the vaccine. In this review, we discuss the rationale for developing DPX-based therapeutic cancer vaccine(s), with a focus on DPX-0907, aimed at inducing efficient anti-tumor immunity that may eventually be shown to prolong patient survival